Microgravity as an Anti-Metastatic Agent in an In Vitro Glioma Model

Maurizio Sabbatini, Valentina Bonetto, Valeria Magnelli, C. Lorusso, Francesco Dondero, Maria Angela Masini
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Abstract

Gravity is a primary physical force that has a profound influence on the stability of the cell cytoskeleton. In our research, we investigated the influence of microgravity on altering the cytoskeletal pathways of glioblastoma cells. The highly infiltrative behavior of glioblastoma is supported by cytoskeletal dynamics and surface proteins that allow glioblastoma cells to avoid stable connections with the tissue environment and other cells. Glioblastoma cell line C6 was exposed to a microgravity environment for 24, 48, and 72 h by 3D-RPM, a laboratory instrument recognized to reproduce the effect of microgravity in cell cultures. The immunofluorescence for GFAP, vinculin, and Connexin-43 was investigated as signals related to cytoskeleton dynamics. The polymerization of GFAP and the expression of focal contact structured by vinculin were found to be altered, especially after 48 and 72 h of microgravity. Connexin-43, involved in several intracellular pathways that critically promote cell motility and invasion of glioma cells, was found to be largely reduced following microgravity exposure. In conclusion, microgravity, by reducing the expression of Connexin-43, alters the architecture of specific cytoskeletal elements such as GFAP and increases the focal contact, which can induce a reduction in glioma cell mobility, thereby inhibiting their aggressive metastatic behavior.
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微重力作为体外胶质瘤模型的抗转移剂
重力是一种主要的物理力,对细胞细胞骨架的稳定性有着深远的影响。在我们的研究中,我们调查了微重力对改变胶质母细胞瘤细胞的细胞骨架通路的影响。胶质母细胞瘤的高度浸润行为得到了细胞骨架动力学和表面蛋白的支持,它们使胶质母细胞瘤细胞避免了与组织环境和其他细胞的稳定连接。胶质母细胞瘤细胞系 C6 通过 3D-RPM 暴露在微重力环境中 24、48 和 72 小时。对 GFAP、vinculin 和 Connexin-43 的免疫荧光进行了研究,这些信号与细胞骨架动力学有关。研究发现,GFAP 的聚合和 vinculin 的焦点接触结构表达发生了改变,尤其是在微重力作用 48 和 72 小时之后。Connexin-43参与了几条细胞内通路,这些通路对促进胶质瘤细胞的运动和侵袭起着至关重要的作用。总之,微重力通过减少Connexin-43的表达,改变了特定细胞骨架元素(如GFAP)的结构,并增加了病灶接触,这可以降低胶质瘤细胞的流动性,从而抑制其侵袭性转移行为。
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